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. 2017 Nov 7;7:14625. doi: 10.1038/s41598-017-15212-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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iPS-RPE cell viability and phenotype following hypoxic stress. (a) Immunohistochemistry images showing RPE cells treated with hypoxyprobe and antibody stained for probe detection. A competed antibody (background staining) was used as control. (b) Images showing iPS-RPE cells stained for HIF2 in normoxia and hypoxia conditions. Nuclei are counterstained with DAPI (blue). Scale bars, 25 μm (c) Western blot analysis showing up-regulation of HIF-1α following 24 hours of hypoxic stress compared to non-stressed cells. N = 5 independent experiments *P < 0,05, t-test. Full-length blots are presented in Supplemental Fig. 3. (d) Representative FACS plots showing the viability of iPS-RPE cells following or not hypoxic stress using the Live/Dead assay. (e) Representative confocal images showing iPS-RPE phenotype following or not hypoxic stress. Cells were stained for: OTX2, Bestrophin, ZO-1, RPE65, EZRIN, CRALBP and Phalloidin. Scale bars, 25 μm.